FI3896171T3 - Menetelmä polynukleotidien prosessoimiseksi yksittäisten solujen analysointia varten - Google Patents
Menetelmä polynukleotidien prosessoimiseksi yksittäisten solujen analysointia varten Download PDFInfo
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- FI3896171T3 FI3896171T3 FIEP21169849.3T FI21169849T FI3896171T3 FI 3896171 T3 FI3896171 T3 FI 3896171T3 FI 21169849 T FI21169849 T FI 21169849T FI 3896171 T3 FI3896171 T3 FI 3896171T3
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- nucleic acid
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- 238000000034 method Methods 0.000 title claims 17
- 102000040430 polynucleotide Human genes 0.000 title 1
- 108091033319 polynucleotide Proteins 0.000 title 1
- 239000002157 polynucleotide Substances 0.000 title 1
- 150000007523 nucleic acids Chemical group 0.000 claims 18
- 108020004707 nucleic acids Proteins 0.000 claims 14
- 102000039446 nucleic acids Human genes 0.000 claims 14
- 239000012491 analyte Substances 0.000 claims 12
- 230000000295 complement effect Effects 0.000 claims 7
- 210000002865 immune cell Anatomy 0.000 claims 6
- 239000002299 complementary DNA Substances 0.000 claims 5
- 108091028043 Nucleic acid sequence Proteins 0.000 claims 4
- 210000004027 cell Anatomy 0.000 claims 4
- 108090000765 processed proteins & peptides Proteins 0.000 claims 4
- 238000010839 reverse transcription Methods 0.000 claims 3
- 229920002477 rna polymer Polymers 0.000 claims 3
- 210000001744 T-lymphocyte Anatomy 0.000 claims 2
- 108020004999 messenger RNA Proteins 0.000 claims 2
- 230000002194 synthesizing effect Effects 0.000 claims 2
- 229920002307 Dextran Polymers 0.000 claims 1
- 108091034117 Oligonucleotide Proteins 0.000 claims 1
- 239000011324 bead Substances 0.000 claims 1
- 210000003850 cellular structure Anatomy 0.000 claims 1
- 239000003153 chemical reaction reagent Substances 0.000 claims 1
- 230000008878 coupling Effects 0.000 claims 1
- 238000010168 coupling process Methods 0.000 claims 1
- 238000005859 coupling reaction Methods 0.000 claims 1
- 230000009089 cytolysis Effects 0.000 claims 1
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 claims 1
- 210000002443 helper t lymphocyte Anatomy 0.000 claims 1
- 210000000581 natural killer T-cell Anatomy 0.000 claims 1
- 210000002501 natural regulatory T cell Anatomy 0.000 claims 1
- 102000004196 processed proteins & peptides Human genes 0.000 claims 1
- 238000012163 sequencing technique Methods 0.000 claims 1
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- C—CHEMISTRY; METALLURGY
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1075—Isolating an individual clone by screening libraries by coupling phenotype to genotype, not provided for in other groups of this subclass
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
- C12Q1/6874—Methods for sequencing involving nucleic acid arrays, e.g. sequencing by hybridisation
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6804—Nucleic acid analysis using immunogens
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
- C12Q1/683—Hybridisation assays for detection of mutation or polymorphism involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
- C12Q2525/10—Modifications characterised by
- C12Q2525/191—Modifications characterised by incorporating an adaptor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2537/00—Reactions characterised by the reaction format or use of a specific feature
- C12Q2537/10—Reactions characterised by the reaction format or use of a specific feature the purpose or use of
- C12Q2537/143—Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2563/00—Nucleic acid detection characterized by the use of physical, structural and functional properties
- C12Q2563/179—Nucleic acid detection characterized by the use of physical, structural and functional properties the label being a nucleic acid
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Analytical Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
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- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Computational Biology (AREA)
- Plant Pathology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Claims (14)
1. Menetelmä yksittäisten solujen analysoimiseksi, joka menetelmä käsittää seuraavat: (a) saadaan aikaan useita osioita, jossa tietty useiden osioiden osio käsittää solun komponentteja, useita viivakoodimolekyylejä kytkettyinä helmeen ja useita analyyttejä, käsittäen seuraavat: (i) ensimmäinen analyytti, joka on: (a) viestinviejä-ribonukleiinihappo, jonka nukleiinihapposekvenssi tai sen komponentti koodaa vähintään immuunisolureseptorin V(D)J-sekvenssin osaa; tai (b) cDNA-molekyyli, joka — käsittää nukleiinihapposekvenssin tai sen komplementin, joka koodaa vähintään immuunisolureseptorin V(D)J-sekvenssin osaa, jolloin cDNA muodostetaan vastaavan MRNA:n käänteistranskriptiolla, käyttäen poly-T:tä sisältävää aluketta; (ii) toinen analyytti, joka on leimausaine, joka kykenee kytkeytymään solupinnan ominaisuuteen, joka on reseptori, jolloin leimausaine käsittää MHC:n, mukaan — (lukien kokonaiset tai osittaiset MHC-peptidit, kytkettyinä oligonukleotidiin, joka käsittää viivakoodisekvenssin, joka identifioi siihen liittyvän MHC:n; jolloin nämä useat viivakoodimolekyylit käsittävät seuraavat: (i) ensimmäinen yksittäinen viivakoodimolekyyli, joka käsittää ensimmäisen nukleiinihappoviivakoodisekvenssin ja kykenee kytkeytymään ensimmäiseen — analyyttiin; ja (ii) toinen yksittäinen viivakoodimolekyyli, joka käsittää nukleiinihappoviivakoodisekvenssin ja kykenee kytkeytymään toiseen analyyttiin; ja (b) suuressa määrässä tai tietyssä osiossa, (i) kytketään ensimmäinen yksittäinen viivakoodimolekyyli ensimmäiseen analyyttiin ja syntetisoidaan ensimmäinen nukleiinihappomolekyyli, joka käsittää vähintään ensimmäisen nukleiinihappoviivakoodisekvenssin tai sen komplementin osan ja ensimmäisen analyytin tai sen komplementin sekvenssin; ja (i) kytketään toinen yksittäinen viivakoodimolekyyli toiseen analyyttiin ja syntetisoidaan toinen nukleiinihappomolekyyli, joka käsittää vähintään toisen nukleiinihappoviivakoodisekvenssin tai sen komplementin osan ja toisen analyytin tai sen komplementin sekvenssin; ja (c) altistetaan ensimmäinen nukleiinihappomolekyyli ja toinen nukleiinihappomolekyyli tai ensimmäisen nukleiinihappomolekyylin ja/tai toisen — nukleiinihappomolekyylin johdannainen sekvensoinnilla ensimmäisen analyytin ja/tai toisen analyytin luonnehtimiseksi.
2. Patenttivaatimuksen 1 mukainen menetelmä, joka käsittää lisäksi ensimmäisen nukleiinihappomolekyylin ja toisen nukleiinihappomolekyylin tai ensimmäisen = nukleiinihappomolekyylin tai toisen nukleiinihappomolekyylin johdannaisen poistamisen tietystä osiosta.
3. Patenttivaatimuksen 1 tai patenttivaatimuksen 2 mukainen menetelmä, jolloin MHC on MHC-peptidimultimeeri tai MHC-peptidi.
4. Patenttivaatimuksen 3 mukainen menetelmä, jolloin MHC-multimeeri valitaan ryhmästä, joka koostuu MHC-tetrameeristä, MHC-pentameeristä ja MHC:llä — merkitystä dekstraanimolekyylistä.
5. Jonkin patenttivaatimuksen 1-4 mukainen menetelmä, jolloin solu on immuunisolu.
6. Patenttivaatimuksen 5 mukainen menetelmä, jolloin immuunisolu on T-solu.
T. Patenttivaatimuksen 6 mukainen menetelmä, jolloin T-solu valitaan — seuraavista koostuvasta ryhmästä: sytotoksiset T-solut, luonnolliset tappaja-T-solut, säätelijä-T-solut ja auttaja-T-solut.
8. Jonkin patenttivaatimuksen 1-7 mukainen menetelmä, jolloin ensimmäinen yksittäinen viivakoodimolekyyli ja/tai toinen yksittäinen viivakoodimolekyyli käsittää yksilöllisen molekyylin tunnistus (UMI) -sekvenssin.
9. Jonkin patenttivaatimuksen 1-8 mukainen menetelmä, jolloin ensimmäinen analyytti on cDNA-molekyyli, joka käsittää nukleiinihapposekvenssin tai sen komplementin, joka koodaa vähintään immuunisolureseptorin V(D)J-sekvenssin osaa, jolloin cDNA muodostetaan vastaavan mRNA:n käänteistranskriptiolla, käyttäen poly-T:tä sisältävää aluketta.
10. Patenttivaatimuksen 9 mukainen menetelmä, jolloin ensimmäinen yksittäinen viivakoodimolekyyli käsittää vaihto-oligon.
11. — Jonkin patenttivaatimuksen 1-8 mukainen menetelmä, jolloin ensimmäinen analyytti on viestinviejä-ribonukleiinihappo, jonka nukleiinihapposekvenssi tai komplementti koodaa vähintään immuunisolureseptorin V(D)J-sekvenssin osaa.
12. Patenttivaatimuksen 11 mukainen menetelmä, jolloin menetelmä käsittää lisäksi cDNA:n muodostamisen viestinviejä-ribonukleiinihapon käänteistranskriptiosta käyttäen templaatin vaihtoa.
13. — Jonkin patenttivaatimuksen 1-12 mukainen menetelmä, jolloin tietty osio — käsittää lisäksi lyysausreagenssia.
14. — Jonkin patenttivaatimuksen 1-13 mukaisen menetelmän käyttö TCR- sekvenssien kokoamiseksi ja kuhunkin soluun liittyvien MHC-peptidiviivakoodien lukumäärän määrittämiseksi.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201662438341P | 2016-12-22 | 2016-12-22 | |
| US15/720,085 US10011872B1 (en) | 2016-12-22 | 2017-09-29 | Methods and systems for processing polynucleotides |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| FI3896171T3 true FI3896171T3 (fi) | 2023-03-19 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| FIEP21169849.3T FI3896171T3 (fi) | 2016-12-22 | 2017-12-22 | Menetelmä polynukleotidien prosessoimiseksi yksittäisten solujen analysointia varten |
Country Status (6)
| Country | Link |
|---|---|
| US (3) | US10011872B1 (fi) |
| EP (5) | EP3559265B1 (fi) |
| CN (2) | CN113801916B (fi) |
| DK (2) | DK3896171T5 (fi) |
| FI (1) | FI3896171T3 (fi) |
| WO (1) | WO2018119447A2 (fi) |
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